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A Battery of Cell- and Structure-specific Markers for the Adult Porcine Retina.

Englund Johansson, Ulrica LU ; Eftekhari, Sajedeh LU and Warfvinge, Karin LU (2010) In The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 58. p.377-389
Abstract
The pig is becoming an increasingly used non-primate model in experimental studies of human retinal diseases and disorders. The anatomy, size, and vasculature of the porcine eye and retina closely resemble the human counterparts, which allows for application of standard instrumentation and diagnostics used in the clinic. Despite the many reports demonstrating immunohistochemistry as a useful method for exploring neuropathological changes in the mammalian central nervous system, including the pig, the porcine retina has been sparsely described. Hence, to facilitate further immunohistochemical analysis of the porcine retina we here report on the successful use of a battery of antibodies for staining of paraformaldehyde fixed cryosectioned... (More)
The pig is becoming an increasingly used non-primate model in experimental studies of human retinal diseases and disorders. The anatomy, size, and vasculature of the porcine eye and retina closely resemble the human counterparts, which allows for application of standard instrumentation and diagnostics used in the clinic. Despite the many reports demonstrating immunohistochemistry as a useful method for exploring neuropathological changes in the mammalian central nervous system, including the pig, the porcine retina has been sparsely described. Hence, to facilitate further immunohistochemical analysis of the porcine retina we here report on the successful use of a battery of antibodies for staining of paraformaldehyde fixed cryosectioned retina. The following antibodies were evaluated for neuronal cells and structures: recoverin (cones and rods), Rho4D2 (rods), transducin-gamma (cones), ROM-1 (photoreceptor outer segments), calbindin (horizontal cells), PKC-alpha (bipolar cells), parvalbumin (amacrine- and displaced amacrine cells), NeuN (ganglion cells, displaced amacrines). For detecting synaptic connections in fibre layers an antibody against synaptobrevin was used. For detecting the retinal pigment epithelium antibodies against cytokeratin and RPE65, respectively, were studied. The glial cell markers used were: bFGF (Müller cells and displaced amacrine cells), GFAP (Müller cells and astrocytes), and vimentin (Müller cells). Each staining was evaluated with regard to its specificity, sensitivity, and reproducibility in the identification of individual cells, specific cell structures and fiber layers, respectively. The markers parvalbumin and ROM-1 were tested here for the first time for the porcine retina. All antibodies tested resulted in specific staining of high quality. In conclusion, all immunohistochemical protocols presented here will be applicable in fixed cryosectioned pig retina. (Less)
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author
organization
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type
Contribution to journal
publication status
published
subject
in
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
volume
58
pages
377 - 389
publisher
Histochemical Society
external identifiers
  • WOS:000276006000008
  • PMID:20086234
  • Scopus:77949786224
ISSN
1551-5044
DOI
10.1369/jhc.2009.954933
language
English
LU publication?
yes
id
e451fc6a-75ed-4167-adfd-67370cc4f248 (old id 1540814)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/20086234?dopt=Abstract
date added to LUP
2010-02-03 09:52:32
date last changed
2017-01-01 07:48:25
@article{e451fc6a-75ed-4167-adfd-67370cc4f248,
  abstract     = {The pig is becoming an increasingly used non-primate model in experimental studies of human retinal diseases and disorders. The anatomy, size, and vasculature of the porcine eye and retina closely resemble the human counterparts, which allows for application of standard instrumentation and diagnostics used in the clinic. Despite the many reports demonstrating immunohistochemistry as a useful method for exploring neuropathological changes in the mammalian central nervous system, including the pig, the porcine retina has been sparsely described. Hence, to facilitate further immunohistochemical analysis of the porcine retina we here report on the successful use of a battery of antibodies for staining of paraformaldehyde fixed cryosectioned retina. The following antibodies were evaluated for neuronal cells and structures: recoverin (cones and rods), Rho4D2 (rods), transducin-gamma (cones), ROM-1 (photoreceptor outer segments), calbindin (horizontal cells), PKC-alpha (bipolar cells), parvalbumin (amacrine- and displaced amacrine cells), NeuN (ganglion cells, displaced amacrines). For detecting synaptic connections in fibre layers an antibody against synaptobrevin was used. For detecting the retinal pigment epithelium antibodies against cytokeratin and RPE65, respectively, were studied. The glial cell markers used were: bFGF (Müller cells and displaced amacrine cells), GFAP (Müller cells and astrocytes), and vimentin (Müller cells). Each staining was evaluated with regard to its specificity, sensitivity, and reproducibility in the identification of individual cells, specific cell structures and fiber layers, respectively. The markers parvalbumin and ROM-1 were tested here for the first time for the porcine retina. All antibodies tested resulted in specific staining of high quality. In conclusion, all immunohistochemical protocols presented here will be applicable in fixed cryosectioned pig retina.},
  author       = {Englund Johansson, Ulrica and Eftekhari, Sajedeh and Warfvinge, Karin},
  issn         = {1551-5044},
  language     = {eng},
  pages        = {377--389},
  publisher    = {Histochemical Society},
  series       = {The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society},
  title        = {A Battery of Cell- and Structure-specific Markers for the Adult Porcine Retina.},
  url          = {http://dx.doi.org/10.1369/jhc.2009.954933},
  volume       = {58},
  year         = {2010},
}